LOCUS       V042682                 7990 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042682
VERSION     V042682
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..2646
                     /label="Plcz1(NM_054066)"
                     /note="Plcz1(NM_054066)"
                     /gene="Plcz1"
     CDS             2653..2742
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2766..3354
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3420..3468
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3670..4098
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4112..4441
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4508..5302
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5478..5599
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5648..5664)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5672..5688
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5696..5726)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5741..5762
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6050..6638)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6809..7669)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7670..7774)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgga aagccaactt
      721 catgagctcg cagaagcaag atggtttttg tcaaaggttc aggatgattt tagaggtgga
      781 aaaatcaacg ttgaaattac tcacaaactg cttgagaaac ttgatttccc atgccacttt
      841 gctcatgtga aacatatttt taaggaaaat gacagacaga accaaggaag aatcaccatt
      901 gaagagttta gagccattta ccggtgtatt gtacatagag aagaaatcac ggagattttc
      961 aacacgtata ctgaaaatag gaaaattctt tctgagaaca gtctgattga gtttctaacc
     1021 caagagcagt atgaaatgga gatcgatcac tctgattcag tagagatcat caataagtat
     1081 gagcccattg aagaagtaaa gggtgagcga cagatgtcaa ttgaaggttt cgcaagatac
     1141 atgttttcat cagaatgtct actgtttaaa gagaactgta aaaccgtgta ccaagatatg
     1201 aatcatccat taagtgatta ttttatttca tcatctcaca acacatattt gatatccgat
     1261 caaatattgg gaccgagtga catttgggga tatgtaagtg ctcttgtgaa aggctgccgc
     1321 tgtctggaaa ttgactgctg ggatggatcc caaaatgaac ccattgtgta ccatggttac
     1381 acattcacca gcaagcttct cttcaaaact gtggtccaag caataaacaa gtatgccttt
     1441 gtgacatctg attacccagt agtgctgtcc ttagaaaatc actgctcccc tggtcagcag
     1501 gaagtgatgg ctagcattct gcagagcacc tttggagact tcctgctttc ggacatgctt
     1561 gaggagtttc cagatacact accgtctcca gaggcactaa aattcaaaat attagtgaaa
     1621 aacaggaaag tgggaacctt atctgaaacc cacgagagga taggaaccga caaaagtggc
     1681 caagtgctag aatggaaaga agtcatctat gaagatggtg atgaagactc aggaatggat
     1741 ccagaaacat gggatgtctt cctatcacgg atcaaggagg agagggaagc agatccctcg
     1801 acattgagtg gaatagcagg cgtcaagaaa aggaagagga agatgaaaat agccatggcc
     1861 ttatctgatc ttgtcattta tactaaggct gagaagttcc gaaacttcca atattcaaga
     1921 gtctatcagc aatttaatga gaccaattcg attggagagt ctcgagctcg aaaactttcc
     1981 aaattgagag tccatgagtt tattttccac accgcggcat tcatcaccag agtatacccc
     2041 aaaatgatga gagcagactc ttctaacttt aaccctcaag agttttggaa tgtaggatgt
     2101 cagatggtgg ccttgaactt tcaaacccct ggactgccta tggatttgca aaacgggaaa
     2161 tttttggata atggaggctc tggatatatt ttgaagccag acatccttag agatacaacc
     2221 ctgggcttta acccaaatga accagaatat gacgaccatc cagttaccct cacaatccga
     2281 atcatcagtg ggatccagtt gcctgttagc tcatcctcta acacgcctga catagtagtg
     2341 atcatagaag tctacggtgt tccaaacgac cacgtgaagc agcagactcg tgttgttaag
     2401 aataatgctt ttagtccaaa gtggaatgaa acatttacat ttcttattca agtgccagaa
     2461 ctggcattga tacgttttgt tgttgaaact caacaaggct tattatcagg aaatgaatta
     2521 ctcgggcagt acactttacc agttctttgc atgaacaaag gttatcgtcg tgttcctctg
     2581 ttttccaaat ccggtgcgaa ccttgaacct tcctcactgt ttatttatgt ttggtacttc
     2641 agagagctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt
     2701 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagtgagg gcccgatatc
     2761 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2821 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2881 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2941 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3001 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3061 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3121 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     3181 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3241 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3301 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     3361 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     3421 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     3481 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     3541 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     3601 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg
     3661 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3721 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3781 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     3841 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3901 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3961 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4021 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     4081 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4141 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4201 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4261 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4321 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4381 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4441 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4501 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4561 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4621 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4681 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4741 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4801 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4861 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4921 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4981 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5041 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5101 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5161 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5221 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5281 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     5341 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5401 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5461 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5521 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5581 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5641 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5701 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5761 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5821 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5881 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5941 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6001 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6061 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6121 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6181 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6241 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6301 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6361 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6421 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6481 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6661 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6721 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6781 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6841 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6901 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6961 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7021 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7081 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7141 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7201 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7261 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7321 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7381 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7441 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7501 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7561 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7621 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7681 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7741 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7801 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc
     7861 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     7921 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     7981 tctgcttagg
//
